Piperazine derivative

ABSTRACT

A piperazine compound represented by formula (1):                    
     wherein X is —CH 2 —, —C(O)— or —CH(CH 3 )—; R 1  is a hydrogen atom or alkyl group; and R 2  is a hydrogen atom, alkyl group, hydroxyalkyl group, arylalkyl group, heteroarylalkyl group, carboxyalkyl group, carboxamidoalkyl group, aminoalkyl group or guanidinoalkyl group; an acid-addition salt thereof, or a hydrate thereof. The compound has excellent inhibitory effects on both cell adhesion and cell infiltration and is useful for prevention or treatment of diseases such as allergy, asthma, rheumatism, arteriosclerosis and inflammation.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to novel cyclic diamine compounds whichhave inhibitory effects on both cell adhesion and cell infiltration andare useful as anti-asthmatic agents, anti-allergic agents,anti-rheumatic agents, anti-arteriosclerotic agents, anti-inflammatoryagents or the like, and medicines containing such compounds.

2. Description of the Background Art

In various inflammatory diseases, infiltration of leukocytes intoinflammatory sites is observed. For example, infiltration of eosinophilsinto the bronchus in asthma (Ohkawara, Y. et al., Am. J. Respir. CellMol. Biol., 12, 4-12 (1995)), infiltration of macrophages and Tlymphocytes into the aorta in arteriosclerosis (Sakai, A. et al.,Arterioscler Thromb. Vasc. Biol., 17, 310-316 (1997)), infiltration of Tlymphocytes and eosinophils into the skin in atopic dermatitis (WakitaH. et al., J. Cutan. Pathol., 21, 33-39 (1994)) or contact dermatitis(Satoh, T. et al., Eur. J. Immunol., 27, 85-91 (1997)), and infiltrationof various leukocytes into rheumatoid synovial tissue (Tak, P P. et al.,Clin. Immunol. Immunopathol., 77, 236-242 (1995)), have been reported.

Infiltration of these leukocytes is elicited by cytokines, chemokines,lipids, and complements produced in inflammatory sites (Albelda, S M. etal., FASEB J., 8, 504-512 (1994)). Activated leukocytes adhere tovascular endothelial cells through an interaction called rolling ortethering with endothelial cells activated likewise. Thereafter, theleukocytes transmigrate through endothelium to infiltrate into theinflammatory sites (Springer, T A., Annu. Rev. Physiol., 57, 827-872(1995)). In the adhesion of leukocytes to the vascular endothelial cellsin this process, various cell adhesion molecules such as animmunoglobulin superfamily (ICAM-1, VCAM-1 and the like), a selectinfamily (E-selectin and the like), an integrin family (LFA-1, VLA-4 andthe like) and CD44, which are induced on the surfaces of the cells bystimulation by cytokines or the like, play important roles (“RinshoMeneki (Clinical Immune)”, 30, Supple. 18 (1998)), and a relationshipbetween the disorder state and aberrant expression of the cell adhesionmolecules is noted.

Accordingly, an agent capable of inhibiting cell adhesion can be usefulas an agent for preventing and treating allergic diseases such asbronchial asthma, dermatitis, rhinitis and conjunctivitis; autoimmunediseases such as rheumatoid arthritis, nephritis, inflammatory boweldiseases, diabetes and arteriosclerosis; and chronic inflammatorydiseases. In fact, it has been reported that antibodies against adhesionmolecules on leukocytes such as LFA-1, Mac-1 and VLA-4 or antibodiesagainst ICAM-1, VCAM-1, P-selectin, E-selectin and the like on vascularendothelial cells, which become ligands thereof, inhibit infiltration ofleukocytes into inflammatory sites in animal models. For example,neutralizing antibodies against VCAM-1 and VLA-4, which is a counterreceptor thereof, can delay development of diabetes in an NOD mousemodel which spontaneously causes the diabetes (Michie, S A. et al.,Curr. Top. Microbiol. Immunol., 231, 65-83 (1998)). It has also beenreported that an antibody against VLA-4 or ICAM-1 and its counterreceptor, LFA-1, inhibits infiltration of eosinophils in a guinea pigand mouse allergic conjunctivitis model (Ebihara et al., Current EyeRes., 19, 20-25 (1999); Whitcup, S M et al., Clin. Immunol., 93, 107-113(1999)), and a monoclonal antibody against VCAM-1 inhibits infiltrationof leukocytes in mouse DSS-induced colitis model to attenuate colitis(Soriano, A. et al., Lab. Invest., 80, 1541-1551 (2000)). Further, ananti-VLA-4 antibody and an anti-CD44 antibody reduce the incidence ofdisease symptoms in a mouse collagen arthritis model (Zeidler, A. etal., Autoimmunity, 21, 245-252 (1995)). Even in cell adhesion moleculedeficient-mice, inhibition of infiltration of leukocytes intoinflammatory tissues is observed likewise in inflammatory models(Bendjelloul, F. et al., Clin. Exp. Immunol., 119, 57-63 (2000);Wolyniec, W W. et al., Am. J. Respir. Cell Mol. Biol., 18, 777-785(1998); Bullard, DC. et al., J. Immunol., 157, 3153-3158 (1996)).

However, it is difficult to develop antibody-based drugs because theyare polypeptides and so oral administration is a problem. Moreover,problems of the possible side effects due to antigenicity and allergicreactions are problems.

On the other hand, there have been various investigations oflow-molecular weight compounds having an inhibitory effect on celladhesion with a view toward permitting oral administration. Thesecompounds include benzothiophene derivatives (Boschelli, D H. et al., J.Med. Chem., 38, 4597-4614 (1995)), naphthalene derivatives (JapanesePatent Application Laid-Open No. 10-147568), hydroxybenzoic acidderivatives (Japanese Patent Application Laid-Open No. 10-182550),lignans (Japanese Patent Application Laid-Open No. 2000-086641 throughPCT route), condensed pyrazine compounds (Japanese Patent ApplicationLaid-Open No. 2000-319277 through PCT route), 2,6-dialkyl-4-silylphenol(Japanese Patent Application Laid-Open Re-Publication No. 2000-509070through PCT route) and the like. However, the goal has not often beensufficiently achieved under the circumstances. Cyclic diamine compoundsdescribed in Japanese Patent Application Laid-Open Nos. 9-143075 and11-92382 do not exhibit a sufficient inhibitory effect on cell adhesion,and there is a demand for further improvement in activity.

SUMMARY OF THE INVENTION

An object of the present invention is to provide a substance havinginhibitory effects on both cell adhesion and cell infiltration, plusexcellent anti-asthmatic effects, anti-allergic effects, anti-rheumaticeffects, anti-arteriosclerotic effects and anti-inflammatory effects.

With the foregoing circumstances in mind, the present inventors carriedout an extensive investigation to find a substance which inhibits celladhesion and cell infiltration. As a result, we found that compoundsrepresented by the general formula (1) have excellent celladhesion-inhibiting effects and cell infiltration-inhibiting effects andare useful as anti-allergic agents, anti-asthmatic agents,anti-rheumatic agents, anti-arteriosclerotic agents or anti-inflammatoryagents.

The present invention provides a piperazine compound represented by thefollowing general formula (1):

wherein X is —CH₂—, —C(O)— or —CH(CH₃)—; R¹ is a hydrogen atom or alkylgroup; and R² is a hydrogen atom, alkyl group, hydroxyalkyl group,arylalkyl group, heteroarylalkyl group, carboxyalkyl group,carboxamidoalkyl group, aminoalkyl group or guanidinoalkyl group;

an acid-addition salt thereof, or a hydrate thereof.

According to the present invention, there is also provided a medicinecomprising the above piperazine compound, the acid-addition saltthereof, or the hydrate thereof as an active ingredient.

According to the present invention, there is further provided amedicinal composition comprising the above piperazine compound, theacid-addition salt thereof, or the hydrate thereof and apharmaceutically acceptable carrier.

According to the present invention, there is still further provided amethod for treating a disease caused by cell adhesion and/or cellinfiltration, which comprises administering an effective amount of theabove piperazine compound, the acid-addition salt thereof, or thehydrate thereof to a patient who requires the treatment.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The alkyl groups represented by R¹ and R² are preferably C₁-C₆-alkylgroups, and specific examples include methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl and hexylgroups, with methyl, ethyl, n-propyl, isopropyl, isobutyl and sec-butylgroups being particularly preferred.

The hydroxyalkyl group represented by R² is a hydroxy-C₁-C₆-alkyl group,and specific examples thereof include hydroxymethyl, 2-hydroxyethyl,2-hydroxy-1-methylethyl, 2-hydroxy-1,1-dimethylethyl, 3-hydroxypropyl,3-hydroxy-2-methylpropyl, 4-hydroxybutyl, 5-hydroxypentyl and6-hydroxyhexyl groups, with hydroxymethyl, 2-hydroxyethyl,2-hydroxy-1-methylethyl, 2-hydroxy-1,1-dimethylethyl and 3-hydroxypropylgroups being particularly preferred. The arylalkyl group is preferably aC₆-C₁₀-aryl-C₁-C₆-alkyl groups, and specific examples thereof includephenyl-C₁-C₆-alkyl groups such as benzyl and phenethyl groups. Theheteroarylalkyl group is preferably a 5- or 6-memberedheteroaryl-C₁-C₆-alkyl group having 1 or 2 nitrogen atoms, andpreferable examples thereof include pyridyl-C₁-C₆-alkyl groups,pyrimidyl-C₁-C₆-alkyl groups, imidazolyl-C₁-C₆-alkyl groups andpyrrolyl-C₁-C₆-alkyl groups. The carboxyalkyl group is preferably acarboxy-C₁-C₆-alkyl group, and specific examples thereof includecarboxymethyl and carboxyethyl groups. The carboxamidoalkyl group is acarboxamido-C₁-C₆-alkyl group, and specific examples thereof includecarboxamidomethyl and carboxamidoethyl. The aminoalkyl group ispreferably a amino-C₁-C₆-alkyl group, and specific examples thereofinclude aminomethyl, aminoethyl and aminopropyl groups. Theguanidinoalkyl group is preferably a guanidino-C₁-C₆-alkyl group, andspecific examples thereof include guanidinomethyl, guanidinoethyl andguanidinopropyl groups.

No particular limitation is imposed on the acid-addition salts of thecompounds (1) according to the invention as long as they arepharmaceutically acceptable salts. Examples include the acid-additionsalts of mineral acids, such as hydrochlorides, hydrobromides,hydriodides, sulfates and phosphates; and acid-addition salts of organicacids, such as benzoates, methanesulfonates, ethanesulfonates,benzenesulfonates, p-toluenesulfonates, oxalates, maleates, fumarates,tartrates, citrates and acetates.

The compounds of formula (1) may be present in the form of solvatestypified by hydrates, and the solvates are embraced in the presentinvention. Optical isomers are present in the compounds (1) according tothe present invention, and all the isomers are also embraced in thepresent invention.

Among the compounds (1) those in which X is —CH(CH₃)— or —CH₂— can beprepared in accordance with, for example, a process shown in thefollowing reaction formula:

wherein X is —CH (CH₃)— or —CH₂—, and R¹ and R² have the same meaningsas defined above.

More specifically, a chlorinated compound (2) is reacted with3,4,5-trimethoxyphenylboronic acid (3) at 0° C. to reflux temperature,preferably 90° C. for 10 minutes to several days, preferably 5 hours inthe presence of a metal catalyst such astetrakis(triphenylphosphine)palladium(0) and a base such as 2 M sodiumcarbonate in a solvent such as toluene, benzene, tetrahydrofuran (THF) ,dioxane or acetonitrile, thereby obtaining a condensate (4). Thiscompound is reacted with lithium aluminum hydride at −20° C. to roomtemperature, preferably at 0° C. for several seconds to several hours,preferably 30 minutes in THF, thereby giving an alcohol (5). Thecompound (5) is stirred together with thionyl chloride at −20° C. toroom temperature, preferably 0° C. for 1 hour to several days,preferably 5 hours in a solvent such as chloroform, dichloromethane,ethyl acetate, ether, THF or dioxane, thereby obtaining achloro-derivative (6). The compound (6) and a diamine (7) are stirred atroom temperature to 100° C., preferably 50° C. for 1 hour to severaldays, preferably 5 hours in the presence of potassium carbonate in asolvent such as N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO)or acetonitrile, thereby obtaining a compound (1a) according to thepresent invention.

Among compounds (1), those in which X is —C(O)— can be prepared inaccordance with, for example, a process shown in the following reactionformula:

wherein X is —C(O)—, and R¹ and R² have the same meanings as definedabove.

More specifically, a glycine methyl ester (8) is reacted with2-nitrobenznesulfonyl chloride in accordance with an already knownmethod, thereby obtaining a 2-nitrobenzenesulofonylated compound (9).The above-described chloro-derivative (6) is reacted with the compound(9) under the same conditions as described above to obtain a compound(10). The compound (10) is treated by an already known method, therebyobtaining a compound (11). The compound (11) is reduced by lithiumaluminum hydride under the same conditions as described above, therebyobtaining an alcohol (12). The compound (12) is reacted withtert-butyldimethylsilyl chloride (TBDMS-Cl) at 0° C. to refluxtemperature, preferably 50° C. for 1 hour to several days, preferably anight in the presence of a base such as imidazole, triethylamine or4-methylmorpholine and 4-(dimethylamino)pyridine in a solventdichloromethane, acetonitrile or DMF to obtain a TBDMS-derivative (13).The compound (13) is reacted with 9-fluorenylmethoxycarbonyl-amino acid(Fmoc-amino acid) (14) at 0° C. to reflux temperature, preferably roomtemperature for 1 minute to several days, preferably 10 minutes in thepresence of a dehydration-condensing agent such asdicyclohexylcarbodiimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimidehydrochloride (water-soluble carbodiimide hydrochloride) orO-(1H-benzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HBTU) in a solvent such as chloroform,dichloromethane, acetonitrile, THF, DMF or DMSO, thereby obtaining acompound (15). The compound (15) is reacted with piperidine inaccordance with an already known method, thereby obtaining an aminederivative (16). The compound (16) is reacted with the above-described2-nitrobenzenesulfonyl chloride under the same conditions as describedabove to obtain a 2-nitrobenzenesulfonylated compound (17). The compound(17) is treated by an already known method, thereby obtaining an alcohol(18). The compound (18) is dissolved in a solvent such as THF or dioxaneand reacted with triphenylphosphine and diethyl azodicarboxylate (DEAD)at 0° C. to reflux temperature, preferably room temperature for 1 hourto several days, preferably a night, thereby obtaining a compound (19).The compound (19) is subjected to de-2-nitrobenzenesulfonylation by analready known method to obtain a compound (20). The above-describedchloro-derivative (6) is reacted with the compound (20) under the sameconditions as described above, thereby obtaining a compound (1b)according to the present invention.

Among the compounds (1) according to the present invention, those inwhich X is —CH₂— can be prepared in accordance with, for example, aprocess shown in the following reaction formula:

wherein X is —CH₂—, and R¹ and R² have the same meanings as definedabove.

More specifically, an N-benzylglycine methyl ester (21) is reacted withFmoc-amino acid (22) in accordance with an already known method, therebyobtaining a dipeptide derivative (23). The compound (23) is subjected tode-Fmoc and cyclization at the same time in accordance with an alreadyknown method to obtain a diketopiperazine derivative (24). The compound(24) is treated by an already known reduction method making use oflithium aluminum hydride or the like, thereby obtaining a piperazinederivative (25). The compound (25) is subjected to de-benzylation byalready known catalytic reduction making use of palladium on carbon,thereby obtaining a compound (26). The compound (26) is reacted with theabove-described chloro-derivative (6) under the same conditions asdescribed above, thereby obtaining a compound (1c) according to thepresent invention.

The compounds (1) according to the present invention are obtained by anyof the above-described processes and may further be purified by using anordinary purification means such as recrystallization or columnchromatography as needed. As needed, the compounds may also be convertedinto the desired salts or solvates in a method known per se in the art.When the compounds (1) include configurational isomers, the presentinvention include any isomers.

The compounds (1) according to the present invention, or acid-additionsalts or solvates thereof thus obtained have an excellent inhibitoryeffect of cell adhesion as demonstrated in the Examples, which will bedescribed subsequently, and are useful as medicines for treatment orprevention of diseases of animals including human, such as asthma,allergy, rheumatism, arteriosclerosis and inflammation.

The medicine according to the present invention comprises a compound(1), a salt thereof, or a solvate thereof as an active ingredient. Theform of administration may be suitably selected as necessary for thetherapeutic application intended without any particular limitation,including oral preparations, injections, suppositories, ointments,inhalants, eye drops, nose drops and plasters. A composition suitablefor use in these administration forms can be prepared by blending apharmaceutically acceptable carrier in accordance with the conventionalpreparation method publicly known by those skilled in the art.

When an oral solid preparation is formulated, an excipient, andoptionally, a binder, a disintegrator, a lubricant, a colorant, a tastecorrigent, a smell corrigent and the like are added to compound (1), andthe resultant composition can be formulated into tablets, coatedtablets, granules, powders, capsules, etc. in accordance with methodsknown in the art.

As such additives described above, any additives may be used which aregenerally used in the pharmaceutical field. Examples thereof includeexcipients such as lactose, sucrose, sodium chloride, glucose, starch,calcium carbonate, kaolin, microcrystalline cellulose and silicic acid;binders such as water, ethanol, propanol, simple syrup, glucosesolution, starch solution, gelatin solution, carboxymethyl cellulose,hydroxypropyl cellulose, hydroxypropyl starch, methyl cellulose, ethylcellulose, shellac, calcium phosphate and polyvinyl pyrrolidone;disintegrators such as dry starch, sodium alginate, agar powder, sodiumhydrogencarbonate, calcium carbonate, sodium lauryl sulfate,monoglyceryl stearate and lactose; lubricants such as purified talc,stearic acid salts, borax and polyethylene glycol; and taste corrigentssuch as sucrose, orange peel, citric acid and tartaric acid.

When an oral liquid preparation is formulated, a taste corrigent,buffer, stabilizer, smell corrigent and/or the like are added tocompound (1), and the resulting composition can be formulated intointernal liquid preparations, syrup preparations, elixirs, etc. inaccordance with methods known in the art. In this case, vanillin as thetaste corrigent, may be used. As the buffer, sodium citrate may bementioned. As examples of the stabilizer, tragacanth, gum arabic andgelatin may be mentioned.

When an injection is formulated, a pH adjustor, buffer, stabilizer,isotonicity agent, local anesthetic and the like may be added to thecompound (1) according to the present invention, and the resultingcomposition can be formulated into subcutaneous, intramuscular andintravenous injections in accordance with methods known in the art.Examples of the pH adjustor and buffer in this case include sodiumcitrate, sodium acetate and sodium phosphate. Examples of the stabilizerinclude sodium pyrosulfite, EDTA, thioglycolic acid and thiolactic acid.Examples of the local anesthetic include procaine hydrochloride andlidocaine hydrochloride. Examples of the isotonicity agent includesodium chloride and glucose.

When a suppository is formulated, a carrier preparation known in theart, for example, polyethylene glycol, lanoline, cacao butter, fattyacid triglyceride or the like, and optionally, a surfactant such asTween (trade mark) and the like are added to the compound (1), and theresultant composition can be formulated into suppositories in accordancewith methods known in the art.

When an ointment is formulated, a base material, stabilizer, wettingagent, preservative and the like, which are generally used, are blendedwith compound (1) as needed, and the resulting blend is mixed andformulated into ointments in accordance with known methods. Examples ofthe base material include liquid paraffin, white vaseline, bleachedbeeswax, octyldodecyl alcohol and paraffin. Examples of the preservativeinclude methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate and propylp-hydroxybenzoate.

Besides the above preparations, inhalants, eye drops and nose drops mayalso be formulated in accordance with known methods.

The dose of the medicine according to the present invention variesaccording to the age, weight and condition of the patient to be treated,the administration method, the number of times of administration, andthe like. It is however preferred that the medicine is generally orallyor parenterally administered at once or in several portions in a dose of1 to 1,000 mg per day in terms of compound (1), for an adult.

The present invention will hereinafter be described in more detail byExamples. However, the present invention is not limited to theseexamples.

PREPARATION EXAMPLE 1 Synthesis of Ethyl2-(3,4,5-trimethoxyphenyl)isonicotinate

3,4,5-Trimethoxyphenylboronic acid (20.64 g) and ethyl2-chloroisonicotinate (19.06 g) were suspended in a mixed solvent oftoluene (200 mL) and THF (100 mL), and to the suspension 2 M sodiumcarbonate (200 mL) and tetrakis(triphenylphosphine)palladium(0) (5.93 g)were added. The mixture was stirred overnight at 90° C. under an argonatmosphere. Ethyl acetate was added to the reaction mixture to separatean organic layer. The organic layer was washed with saturated brine,dried over anhydrous magnesium sulfate and concentrated under reducedpressure. The residue was purified by column chromatography on silicagel (hexane:ethyl acetate=5:1) to obtain the title compound.

Yield: 27.70 g (85%).

¹H-NMR (400 MHz, CDCl₃) δ: 1.45(t,3H,J=7.0 Hz), 3.92(s,3H), 3.99(s,6H),4.46(q,2H,J=7.0 Hz), 7.30(s,2H), 7.76(dd,1H,J=5.1 Hz,1.6 Hz),8.24(dd,1H,J=1.6 Hz,0.8 Hz), 8.81(dd,1H,J=5.1 Hz,0.8 Hz).

PREPARATION EXAMPLE 2 Synthesis of4-hydroxymethyl-2-(3,4,5-trimethoxyphenyl)-pyridine

Ethyl 2-(3,4,5-trimethoxyphenyl)nicotinate (27.70 g) was dissolved inTHF (200 mL), and to the solution lithium aluminum hydride (3.31 g) wasadded at 0° C. under an argon atmosphere, and the mixture was stirred at0° C. for 1 hour as it is. A small amount of water and then sodiumsulfate were added to the reaction mixture, and the reaction mixture wasfiltered through celite. The filtrate was concentrated under reducedpressure, and the resultant crystals were recrystallized from ethylacetate-hexane to obtain the title compound.

Yield: 18.15 g (76%).

¹H-NMR (400 MHz, CDCl₃) δ: 3.90(s,3H), 3.95(s,6H), 4.79(s,2H),7.19(d,1H,J=5.1 Hz), 7.21(s,2H), 7.66(s,1H), 8.60(d,1H,J=5.1 Hz).

PREPARATION EXAMPLE 3 Synthesis of4-chloromethyl-2-(3,4,5-trimethoxyphenyl)-pyridine:

4-Hydroxymethyl-2-(3,4,5-trimethoxyphenyl)pyridine (18.15 g) wasdissolved in chloroform (300 mL) , and to the solution thionyl chloride(19.2 mL) was added at 0° C.. After 30 minutes, the mixture was warmedto room temperature and stirred for 4 hours. The reaction mixture waswashed with water and saturated brine, dried over anhydrous sodiumsulfate and concentrated under reduced pressure. The resultant crystalswere then recrystallized from chloroform-hexane to obtain the titlecompound.

Yield: 17.87 g (92%).

¹H-NMR (400 MHz, CDCl₃) δ: 3.91(s,3H), 3.97(s,6H) , 4.61 (s,2H) ,7.24(s,2H), 7.26(d,1H,J=5.1 Hz), 7.68(s,1H), 8.67(d,1H,J=5.1 Hz).

EXAMPLE 1 Synthesis ofcis-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-2,6-dimethylpiperazinetetrahydrochloride

4-Chloromethyl-2-(3,4,5-trimethoxyphenyl)pyridine (123 mg) andcis-2,6-dimethylpiperazine (23 mg) were dissolved in DMF (5 mL), and tothe solution potassium carbonate (58 mg) was added. The mixture wasstirred at 80° C. for 4 hours and concentrated under reduced pressure.Water was added to the residue to conduct extraction with chloroform.The resultant organic layer was washed with saturated brine, dried overanhydrous magnesium sulfate and then concentrated under reducedpressure. The residue was purified by column chromatography on silicagel (chloroform:methanol=40:1) to obtain a free base of the titlecompound. This compound was dissolved in ethyl acetate, and to thesolution an ethyl acetate solution of 4 M hydrogen chloride was added toprovide a hydrochloride.

Yield: 101 mg (68%).

¹H-NMR (measured as a free base, 400 MHz, CDCl₃) δ: 0.97(d,6H,J=6.1 Hz),1.99(t,2H,J=11.1 Hz), 2.75(d,4H,J=9.8 Hz), 3.53(s,2H), 3.81(s,2H),3.90(s,6H), 3.97(s,6H),3.98(s,6H), 7.22-7.24(m,5H), 7.32(d,1H,J=4.3 Hz),7.64(s,1H), 7.67(s,1H), 8.57(d,1H,J=5.1 Hz), 8.61(d,1H,J=5.1 Hz). m/z(EI): 628 [M⁺].

EXAMPLE 2 Synthesis oftrans-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]-2,5-dimethylpiperazinetetrahydrochloride

4-Chloromethyl-2-(3,4,5-trimethoxyphenyl)pyridine (123 mg) andtrans-2,5-dimethylpiperazine (23 mg) were reacted in the same manner asin Example 1 to obtain the title compound as a hydrochloride.

Yield: 117 mg (93%).

¹H-NMR (measured as a free base, 400 MHz, CDCl₃) δ: 1.07(d,6H,J=6.1 Hz),2.02(t,2H,J=10.5 Hz), 2.46-2.49(m,2H), 2.67(dd,2H,J=11.2 Hz,2.6 Hz),3.16(d,2H,J=14.4 Hz), 3.91(s,6H), 3.97(s,12H), 4.10(d,2H,J=14.3 Hz),7.24(s,4H), 7.26(d,2H,J=5.3 Hz), 7.63(s,2H), 8.60(d,2H,J=5.1 Hz). m/z(EI): 628 [M⁺].

EXAMPLE 3 Synthesis ofN,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-2-hydroxymethylpiperazinedimaleate:

4-Chloromethyl-2-(3,4,5-trimethoxyphenyl)pyridine (1.44 g) and2-hydroxymethylpiperazine (463 mg) were reacted in the same manner as inExample 1 to obtain the title compound as a maleate.

Yield: 116 mg (19%).

¹H-NMR (measured as a maleate, 400 MHz, DMSO-d₆) δ: 2.37-2.79(m,7H),3.57-3.62(m,4H), 3.78(s,6H), 3.89(s,12H), 4.05-4.11(m,2H), 6.64(s,4H),7.23(d,1H,J=5.1 Hz), 7.26(d,1H,J=4.6 Hz), 7.34(s,2H), 7.34(s,2H),7.76(s,1H), 7.78(s,1H), 8.52(d,1H,J=6.3 Hz), 8.53(d,1H,J=5.4 Hz). m/z(EI): 630 [M⁺].

PREPARATION EXAMPLE 4 Synthesis of N-(2-nitrobenzenesulfonyl)glycinemethyl ester

Glycine methyl ester hydrochloride (15.0 g) was dissolved indichloromethane, and to the solution triethylamine (26.48 g) was addedat 0° C. A solution of 2-nitrobenzenesulfonyl chloride (23.57 g) indichloromethane (50 mL) was then gradually added dropwise. After themixture was stirred at room temperature for 2 hours, the reactionmixture was concentrated under reduced pressure, and ethyl acetate wasadded to the residue. The resultant mixture was washed with 2 Mhydrochloric acid, water and saturated brine, dried over anhydroussodium sulfate and concentrated under reduced pressure. The resultantcrystals were recrystallized from ethyl acetate-hexane to obtain thetitle compound.

Yield: 26.20 g (90%).

¹H-NMR (400 MHz, CDCl₃) δ: 3.61(s,3H), 4.02(d,2H,J=5.9 Hz), 6.07(br,1H),7.73-7.77(m,2H), 7.92-7.95(m,1H), 8.07-8.11(m,1H).

PREPARATION EXAMPLE 5 Synthesis ofN-(2-nitrobenzenesulfonyl)-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]glycinemethyl ester

N-(2-Nitrobenzenesulfonyl)glycine methyl ester (5.60 g) was dissolved inacetonitrile (100 mL), and to the solution potassium carbonate (3.10 g)and potassium iodide (2.29 g) were added. To the mixture,4-chloromethyl-2-(3,4,5-trimethoxyphenyl)pyridine (6.00 g) was thenadded, and the resultant mixture was stirred at 80° C. for 1 hour. Thereaction mixture was concentrated under reduced pressure, ethyl acetatewas added to the residue, and the resultant mixture was washed with asaturated aqueous solution of sodium hydrogencarbonate, water andsaturated brine, dried over anhydrous sodium sulfate and concentratedunder reduced pressure. The residue was purified by columnchromatography on silica gel (chloroform:methanol=50:1) to obtain thetitle compound.

Yield: 11.35 g (theoretical amount).

¹H-NMR (400 MHz, CDCl₃) δ: 3.63(s,3H), 3.90(s,3H), 3.97(s,6H),4.13(s,2H), 4.75(s,2H), 7.13(d,1H,J=3.5 Hz), 7.20(s,2H), 7.60(s,1H),7.65-7.73(m,3H), 8.07(dd,1H,J=8.8 Hz,1.6 Hz), 8.61(d,1H,J=5.1 Hz)

PREPARATION EXAMPLE 6 Synthesis ofN-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]glycine methyl ester

N-(2-Nitrobenzenesulfonyl)-N-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]glycinemethyl ester (11.35 g) was dissolved in acetonitrile (30 mL), and to thesolution potassium carbonate (3.39 g) was added. Thiophenol (2.37 g) wasthen added to the mixture, and the resultant mixture was stirredovernight at room temperature. Ethyl acetate was added to the reactionmixture, and the resultant mixture was washed with a saturated aqueoussolution of sodium hydrogencarbonate, water and saturated brine, driedover anhydrous sodium sulfate and concentrated under reduced pressure.The residue was then purified by column chromatography on silica gel(chloroform:methanol=20:1) to obtain the title compound.

Yield: 6.54 g (92%).

¹H-NMR (400 MHz, CDCl₃) δ: 3.46(s,2H), 3.74(s,3H), 3.90(s,5H),3.97(s,6H), 7.24(s,2H), 7.25(d,1H,J=4.1 Hz), 7.67(s,1H), 8.65(d,1H,J=4.9Hz).

PREPARATION EXAMPLE 7 Synthesis ofN-(2-hydroxyethyl)-N-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]amine:

N-[[2-(3,4,5-Trimethoxyphenyl)pyridin-4-yl]methyl]-glycine methyl ester(6.54 g) was dissolved in THF (80 mL), and to the solution lithiumaluminum hydride (717 mg) was gradually added portionwise at 0° C. underan argon atmosphere, and the mixture was stirred for 4 hours. A smallamount of water was added to the reaction mixture. When bubbling ended,sodium sulfate was excessively added. The reaction mixture was filteredthrough celite, and the filtrate was concentrated under reducedpressure, and the residue was purified by column chromatography onsilica gel (chloroform:methanol=20:1) to obtain the title compound.

Yield: 5.03 g (84%).

¹H-NMR (400 MHz, CDCl₃) δ: 2.14(br,2H), 2.83(t,2H,J=5.1 Hz),3.71(t,2H,J=5.1 Hz), 3.89(s,2H), 3.90(s,3H), 3.96(s,6H), 7.19(d,1H,J=4.9Hz), 7.23(s,2H), 7.64(s,1H), 8.60(d,1H,J=5.1 Hz).

PREPARATION EXAMPLE 8 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]amine

N-(2-Hydroxyethyl)-N-[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]amine(5.0 g) was dissolved in acetonitrile (100 mL), and to the solutiontriethylamine (2.22 g) and 4-(dimethylamino)pyridine (250 mg) wereadded, and tert-butylchlorodimethylsilane (3.08 g) was then added. Themixture was stirred at 50° C. for 4 hours. The reaction, the reactionmixture was concentrated under reduced pressure, ethyl acetate was addedto the residue, and the mixture was washed with water and saturatedbrine, dried over anhydrous sodium sulfate and concentrated underreduced pressure. The residue was purified by column chromatography onsilica gel (chloroform:methanol=30:1) to obtain the title compound.

Yield: 6.89 g (theoretical amount)

¹H-NMR (400 MHz, CDCl₃) δ: 0.07(s,6H), 0.90(s,9H), 1.93(br,1H),2.76(t,2H,J=5.1 Hz), 3.77(t,2H,J=5.1 Hz), 3.90(s,5H), 3.97(s,6H),7.21(d,1H,J=4.7 Hz), 7.24(s,2H), 7.66(s,1H), 8.60(d,1H,J=4.9 Hz).

PREPARATION EXAMPLE 9 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(9-fluorenylmethoxycarbonyl)glycineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]amine(3.40 g), N-(9-fluorenylmethoxycarbonyl)glycine (2.34 g),diisopropylethylamine (1.03 g) and 4-(dimethylamino)pyridine (961 mg)were dissolved in acetonitrile (40 mL), and to the solution HBTU (3.13g) was added, and the mixture was stirred at room temperature for 10minutes. The reaction mixture was concentrated under reduced pressure,ethyl acetate was added to the residue, and the mixture was washed withwater and saturated brine, dried over anhydrous sodium sulfate and thenconcentrated under reduced pressure. The residue was purified by columnchromatography on silica gel (n-hexane:ethyl acetate=1:1) to obtain thetitle compound.

Yield: 5.15 g (92%).

¹H-NMR (400 MHz, CDCl₃) δ: 0.04(s,6H), 0.87(s,9H), 3.43(t,2H,J=5.1 Hz),3.75(t,2H,J=5.1 Hz), 3.90(s,3H), 3.95(s,6H), 4.27(d,1H,J=4.5 Hz),4.34-4.39(m,3H), 4.75(s,2H), 5.83(br,1H), 7.09(d,1H,J=4.1 Hz),7.19(s,2H), 7.30(t,2H,J=7.4 Hz), 7.39(t,2H,J=7.4 Hz), 7.58(s,1H),7.61(d,2H,J=7.6 Hz), 7.76(d,2H,J=7.6 Hz), 8.61(d,1H,J=5.1 Hz).

PREPARATION EXAMPLE 10 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]glycineamide

N-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(9-fluorenylmethoxycarbonyl)glycineamide (5.15 g) was dissolved in a 20% acetonitrile solution (40 mL) ofpiperidine, and the solution was stirred at room temperature for 4hours. Ethyl acetate was added to the reaction mixture, and the mixturewas washed with water and saturated brine, dried over anhydrous sodiumsulfate and then concentrated under reduced pressure. The residue waspurified by column chromatography on silica gel(chloroform:methanol=20:1) to obtain the title compound.

Yield: 2.76 g (78%).

¹H-NMR (400 MHz, CDCl₃) δ: 0.04(s,6H), 0.88(s,9H), 1.67 (br,2H),3.38(t,2H,J=5.2 Hz), 3.70(s,2H), 3.72(t,2H,J=5.2 Hz), 3.90(s,3H),3.96(s,6H), 4.73(s,2H), 7.08(d,1H,J=4.1 Hz), 7.20(s,2H), 7.50(s,1H),8.60(d,1H,J=5.1 Hz).

PREPARATION EXAMPLE 11 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)glycineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]glycineamide (2.52 g) was treated in the same manner as in Preparation Example4 to obtain the title compound.

Yield: 3.41 g (98%).

¹H-NMR (400 MHz, CDCl₃) δ: 0.00(s,6H), 0.83(s,9H), 3.39(t,2H,J=4.8 Hz),3.47(d,2H,J=7.0 Hz), 3.70(t,2H,J=4.8 Hz), 3.89(s,3H), 3.95(s,6H),4.60(s,2H), 6.50(br,1H), 6.93(d,1H,J=4.9 Hz), 7.18(s,2H), 7.44(s,1H),7.61-7.67(m,2H), 7.81(dd,1H,J=7.5 Hz,1.7 Hz), 8.05(dd,1H,J=7.7 Hz,2.0Hz), 8.54(d,1H,J=4.1 Hz).

PREPARATION EXAMPLE 12 Synthesis ofN-[2-hydroxyethyl]-N-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-glycineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)glycineamide (3.41 g) was dissolved in THF, and to the solution a THF solution(6.1 mL) of 1.0 M tetrabutylammonium fluoride was added, and the mixturewas stirred at room temperature for 4 hours. The reaction mixture wasconcentrated under reduced pressure, and ethyl acetate was added to theresidue. The mixture was washed with water and saturated brine, driedover anhydrous sodium sulfate and then concentrated under reducedpressure. The residue was purified by column chromatography on silicagel (chloroform:methanol=10:1) to obtain the title compound.

Yield: 2.22 g (78%).

¹H-NMR (400 MHz, CDCl₃) δ: 3.38(br,2H), 3.55(br,2H), 3.71(br,2H),3.88(s,3H), 3.93(s,6H), 4.56(s,2H), 6.89(d,1H,J=4.9 Hz), 7.19(s,2H),7.46(s,1H), 7.50-7.63(m,2H), 7.78(d,1H,J=7.4 Hz), 8.04(d,1H,J=7.4 Hz),8.49(d,1H,J=4.7 Hz).

PREPARATION EXAMPLE 13 Synthesis of1-(2-nitrobenzenesulfonyl)-3-oxo-4-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine

N-(2-Hydroxyethyl)-N-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-glycineamide (2.22 g) was dissolved in THF, and to the solutiontriphenylphosphine (1.55 g) was added. DEAD (1.03 g) was slowly added tothe mixture at room temperature, and the resultant mixture was stirredovernight at room temperature under an argon atmosphere. After thereaction mixture was concentrated under reduced pressure, ethyl acetatewas added to the residue. The mixture was washed with a saturatedaqueous solution of sodium hydrogencarbonate, water and saturated brine,dried over anhydrous sodium sulfate and then concentrated under reducedpressure. The residue was purified by column chromatography on silicagel (ethyl acetate) to obtain the title compound.

Yield: 2.01 g (94%).

¹H-NMR (400 MHz, CDCl₃) δ: 3.42(t,2H,J=5.2 Hz), 3.67(t,2H,J=5.2 Hz),3.90(s,3H), 3.96(s,6H), 4.07(s,2H), 4.67(s,2H), 7.05(d,1H,J=4.6 Hz),7.20(s,2H), 7.51(s,1H), 7.63(d,1H,J=2.0 Hz), 7.69-7.76(m,2H),8.03(d,1H,J=2.2 Hz), 8.61(d,1H,J=5.1 Hz).

PREPARATION EXAMPLE 14 Synthesis of2-oxo-1-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine

1-(2-Nitrobenzenesulfonyl)-3-oxo-4-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine(1.83 g) was treated in the same manner as in Preparation Example 6 toobtain the title compound.

Yield: 118 mg (10%).

¹H-NMR (400 MHz, CDCl₃) δ: 1.82(br,1H), 3.09(t,2H,J=5.4 Hz),3.29(t,2H,J=5.4 Hz), 3.65(s,2H), 3.90(s,6H), 3.96(s,3H), 4.67(s,2H),7.12(d,1H,J=4.9 Hz), 7.21(s,2H), 7.55(s,1H), 8.63(d,1H,J=5.1 Hz).

EXAMPLE 4 Synthesis of2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]piperazine2.5 hydrochloride

2-Oxo-1-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine (62mg) was dissolved in acetonitrile (5 mL), and to the solution potassiumcarbonate (24 mg), potassium iodide (29 mg) and4-chloromethyl-2-(3,4,5-trimethoxyphenyl)pyridine (51 mg) were added,and the mixture was stirred at 80° C. for 1 hour. The reaction mixturewas concentrated under reduced pressure, ethyl acetate was added to theresidue, and the resultant mixture was washed with a saturated aqueoussolution of sodium hydrogencarbonate, water and saturated brine, driedover anhydrous sodium sulfate and concentrated under reduced pressure.The resultant oil was purified by preparative TLC on silica gel(chloroform:methanol=25:1) to obtain the title compound as ahydrochloride.

Yield: 92 mg (87%).

¹H-NMR (measured as a free base, 400 MHz, CDCl₃) δ: 2.73(t,2H,J=5.1 Hz),3.32(t,2H,J=5.1 Hz), 3.34(s,2H), 3.65(s,2H), 3.90(s,6H), 3.96(s,12H),4.67(s,2H), 7.11(d,1H,J=4.9 Hz), 7.22(br,5H), 7.55(s,1H), 7.61(s,1H),8.63-8.64(m,2H).

m/z (EI): 614 [M⁺].

PREPARATION EXAMPLE 15 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(9-fluorenylmethoxycarbonyl)-L-alanineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]amine(756 mg) and N-(9-fluorenylmethoxycarbonyl)-L-alanine (544 mg) weretreated in the same manner as in Preparation Example 9 to obtain thetitle compound. Since this compound was unable to be isolated fromimpurities, it was used in the next reaction without purifying it as itis.

PREPARATION EXAMPLE 16 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-L-alanineamide

The whole amount of the reaction mixture obtained in Preparation Example15 was treated in the same manner as in Preparation Example 10 to obtainthe title compound.

Yield: 340 mg (39% by 2 steps).

PREPARATION EXAMPLE 17 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-alanineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-L-alanineamide (340 mg) was treated in the same manner as in Preparation Example4 to obtain the title compound.

Yield: 303 mg (65%).

PREPARATION EXAMPLE 18 Synthesis ofN-(2-hydroxyethyl)-N-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-alanineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-alanineamide (669 mg) was treated in the same manner as in Preparation Example12 to obtain the title compound.

Yield: 546 mg (98%).

PREPARATION EXAMPLE 19 Synthesis of(3S)-3-methyl-4-(2-nitrobenzenesulfonyl)-2-oxo-1-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine

N-(2-Hydroxyethyl)-N-[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-alanineamide (546 mg) was treated in the same manner as in Preparation Example13 to obtain the title compound. Since removal of by-products could notbe completely conducted, this compound was used in the next reactionwithout conducting further purification.

PREPARATION EXAMPLE 20 Synthesis of(3S)-3-methyl-2-oxo-1-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]piperazine

The whole amount of the reaction mixture obtained in Preparation Example19 was treated in the same manner as in Preparation Example 6 to obtainthe title compound.

Yield: 174 mg (50% by 2 steps).

¹H-NMR (400 MHz, CDCl₃) δ: 1.47(d,3H,J=6.8 Hz), 1.78(br,1H),3.02-3.09(m,1H), 3.15-3.22(m,2H), 3.39-3.45(m,1H), 3.65(q,1H,J=6.8 Hz),3.90(s,3H), 3.96(s,6H), 4.60(d,1H,J=15.2 Hz), 4.70(d,1H,J=15.2 Hz),7.10(dd,1H,J=5.0 Hz,1.5 Hz), 7.22(s,2H), 7.53(s,1H), 8.62(d,1H,J=4.9Hz).

EXAMPLE 5 Synthesis of(3S)-3-methyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine2.5 hydrochloride

(3S)-3-Methyl-2-oxo-1-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]piperazine(80 mg) and 4-chloromethyl-2-(3,4,5-trimethoxyphenyl)pyridine (63 mg)were reacted in the same manner as in Example 4 to obtain the titlecompound as a hydrochloride.

Yield: 124 mg (92%).

¹H-NMR (measured as a free base, 400 MHz, CDCl₃) δ: 1.56(d,3H,J=6.6 Hz),2.53-2.59(m,1H), 2.95-2.99(m,1H), 3.21-3.33(m,2H), 3.43(q,1H,J=6.8 Hz),3.52(d,1H,J=14.4 Hz), 3.90(s,6H), 3.95(s,6H), 3.95(s,7H),4.58(d,1H,J=15.4 Hz), 4.75(d,1H,J=15.2 Hz), 7.10(d,1H,J=4.7 Hz),7.22(s,2H), 7.23(m,3H), 7.54(s,1H), 7.62(s,1H), 8.61(d,1H,J=5.7 Hz),8.63(d,1H,J=5.9 Hz).

m/z (EI): 628 [M⁺].

PREPARATION EXAMPLE 21 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(9-fluorenylmethoxycarbonyl)-L-valineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]amine(679 mg) and N-(9-fluorenylmethoxycarbonyl)-L-valine (865 mg) weretreated in the same manner as in Preparation Example 9 to obtain thetitle compound.

Yield: 1.11 g (74%).

PREPARATION EXAMPLE 22 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-L-valineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(9-fluorenylmethoxycarbonyl)-L-valineamide (1.11 g) was treated in the same manner as in Preparation Example10 to obtain the title compound.

Yield: 705 mg (90%).

PREPARATION EXAMPLE 23 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-valineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-L-valineamide (705 mg) was treated in the same manner as in Preparation Example4 to obtain the title compound.

Yield: 877 mg (92%).

PREPARATION EXAMPLE 24 Synthesis ofN-(2-hydroxyethyl)-N-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-valineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-valineamide (877 mg) was treated in the same manner as in Preparation Example12 to obtain the title compound.

Yield: 689 mg (95%).

PREPARATION EXAMPLE 25 Synthesis of(3S)-3-isopropyl-4-(2-nitrobenzenesulfonyl)-2-oxo-1-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-piperazine

N-(2-Hydroxyethyl)-N-[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-valineamide (546 mg) was treated in the same manner as in Preparation Example13 to obtain the title compound. Since removal of by-products could notbe completely conducted, this compound was used in the next reactionwithout conducting further purification.

PREPARATION EXAMPLE 26 Synthesis of(3S)-3-isopropyl-2-oxo-1-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine

The whole amount of the reaction mixture obtained in Preparation Example25 was treated in the same manner as in Preparation Example 6 to obtainthe title compound.

Yield: 365 mg (79% by 2 steps).

¹H-NMR (400 MHz, CDCl₃) δ: 0.97(d,3H,J=6.8 Hz), 1.05(d,3H,J=7.0 Hz),1.63(br,1H), 2.56-2.64(m,1H), 2.99-3.31(m,3H), 3.41-3.47(m,2H),3.90(s,3H), 3.95(s,6H), 4.48(d,1H,J=15.4 Hz), 4.90(d,1H,J=15.4 Hz),7.10(dd,1H,J=4.9 Hz,1.2 Hz), 7.21(s,2H), 7.53(s,1H), 8.61(d,1H,J=5.1Hz).

EXAMPLE 6 Synthesis of(3S)-3-isopropyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazinesesqui-hydrochloride

(3S)-3-Isopropyl-2-oxo-1-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]piperazine(80 mg) and 4-chloromethyl-2-(3,4,5-trimethoxyphenyl)pyridine (59 mg)were reacted in the same manner as in Example 4 to obtain the titlecompound as a hydrochloride.

Yield: 98 mg (75%).

¹H-NMR (measured as a free base, 400 MHz, CDCl₃) δ: 1.13(d,3H,J=6.8 Hz),1.22(d,3H,J=6.8 Hz), 2.23-2.27(m,1H), 2.60-2.64(m,1H), 3.03-3.35(m,4H),3.65(d,1H,J=14.8 Hz), 3.90(s,3H), 3.91(s,3H), 3.95(s,6H), 3.96(s,6H),3.96(d,1H,J=14.8 Hz), 4.48(d,1H,J=15.2 Hz), 4.92(d,1H,J=15.2 Hz),7.12(d,1H,J=4.9 Hz), 7.21-7.24(m,5H), 7.56(s,1H), 7.65(s,1H),8.60(d,1H,J=4.9 Hz), 8.63(d,1H,J=5.1 Hz).

m/z (EI): 656 [M⁺].

PREPARATION EXAMPLE 27 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(9-fluorenylmethoxycarbonyl)-D-valineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]amine(1.27 g) and N-(9-fluorenylmethoxycarbonyl)-D-valine (1.00 g) weretreated in the same manner as in Preparation Example 9 to obtain thetitle compound. Since impurities were unable to be removed from thiscompound, the compound was provided to the next step without purifyingit.

PREPARATION EXAMPLE 28 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-D-valineamide

CrudeN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(9-fluorenylmethoxycarbonyl)-D-valineamide obtained in Preparation Example 27 was treated in the same manneras in Preparation Example 10 to obtain the title compound.

Yield: 1.00 g (64% by 2 steps).

PREPARATION EXAMPLE 29 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-D-valineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-D-valineamide (1.00 g) was treated in the same manner as in Preparation Example4 to obtain the title compound.

Yield: 1.36 g (94%).

PREPARATION EXAMPLE 30 Synthesis ofN-(2-hydroxyethyl)-N-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-D-valineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-D-valineamide (1.36 g) was treated in the same manner as in Preparation Example12 to obtain the title compound.

Yield: 1.08 g (94%).

PREPARATION EXAMPLE 31 Synthesis of(2R)-2-isopropyl-1-(2-nitrobenzenesulfonyl)-3-oxo-4-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-piperazine

N-(2-Hydroxyethyl)-N-[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-D-valineamide (258 mg) was treated in the same manner as in Preparation Example13 to obtain the title compound. Since removal of by-products could notbe completely conducted, this compound was used in the next reactionwithout conducting further purification.

PREPARATION EXAMPLE 32 Synthesis of(3R)-3-isopropyl-2-oxo-1-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine

The whole amount of the reaction mixture obtained in Preparation Example31 was treated in the same manner as in Preparation Example 6 to obtainthe title compound.

Yield: 109 mg (63% by 2 steps).

¹H-NMR (400 MHz, CDCl₃) δ: 1.01(d,3H, J=6.6 Hz), 1.08(d,3H,J=7.0 Hz),2.58-2.70(m,1H), 3.05-3.30(m,3H), 3.45-3.57(m,2H), 3.93(s,3H),3.98(s,6H), 4.51(d,1H,J=15.4 Hz), 4.92(d,1H,J=15.4 Hz), 7.12(d,1H,J=5.1Hz), 7.24(s,2H), 7.56(s,1H), 8.64(d,1H,J=4.9 Hz).

EXAMPLE 7 Synthesis of(3R)-3-isopropyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazinetri-hydrochloride

(3R)-3-Isopropyl-2-oxo-1-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]piperazine(109 mg) and 4-chloromethyl-2-(3,4,5-trimethoxyphenyl)pyridine (104 mg)were reacted in the same manner as in Example 4 to obtain the titlecompound as a hydrochloride.

Yield: 58 mg (50%).

¹H-NMR (measured as a free base, 400 MHz, CDCl₁ ₃) δ: 1.14(d,3H,J=7.0Hz), 1.23(d,3H,J=6.8 Hz), 2.20-2.35(m,1H), 2.58-2.70(m,1H),3.03-3.45(m,4H), 3.67(d,1H,J=14.8 Hz), 3.90-3.91(m,6H),3.96-3.97(m,15H), 7.13-7.15(m,1H), 7.24(s,5H), 7.59(s,1H), 7.67(s,1H),8.61(d,1H,J=5.1 Hz), 8.64(d,1H,J=5.1 Hz).

m/z (EI): 656 [M⁺].

PREPARATION EXAMPLE 33 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(9-fluorenylmethoxycarbonyl)-L-leucineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]amine(1.40 g) and N-(9-fluorenylmethoxycarbonyl)-L-leucine (1.16 g) weretreated in the same manner as in Preparation Example 9 to obtain thetitle compound.

Yield: 2.32 g (93%).

PREPARATION EXAMPLE 34 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-L-leucineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(9-fluorenylmethoxycarbonyl)-L-leucineamide (2.32 g) was treated in the same manner as in Preparation Example10 to obtain the title compound.

Yield: 1.57 g (96%).

PREPARATION EXAMPLE 35 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-leucineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-L-leucineamide (1.57 g) was treated in the same manner as in Preparation Example4 to obtain the title compound.

Yield: 2.05 g (98%).

PREPARATION EXAMPLE 36 Synthesis ofN-(2-hydroxyethyl)-N-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-leucineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-leucineamide (2.05 g) was treated in the same manner as in Preparation Example12 to obtain the title compound.

Yield: 1.61 g (93%).

PREPARATION EXAMPLE 37 Synthesis of(3S)-3-(2-methylpropyl)-4-(2-nitrobenzene-sulfonyl)-2-oxo-1-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine

N-(2-Hydroxyethyl)-N-[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-leucineamide (1.57 g) was treated in the same manner as in Preparation Example13 to obtain the title compound. Since removal of by-products could notbe completely conducted, this compound was used in the next reactionwithout conducting further purification.

PREPARATION EXAMPLE 38 Synthesis of(3S)-3-(2-methylpropyl)-2-oxo-1-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine

The whole amount of the reaction mixture obtained in Preparation Example37 was treated in the same manner as in Preparation Example 6 to obtainthe title compound.

Yield: 689 mg (44% by 2 steps).

¹H-NMR (400 MHz, CDCl₃) δ: 0.95(d,3H,J=6.6 Hz), 0.98(d,3H,J=6.6 Hz),1.60(ddd,1H,J=13.7 Hz,9.9 Hz,4.2 Hz), 1.71(br,1H), 1.77-1.80(m,1H),1.95(ddd,1H,J=13.7 Hz,9.9 Hz,4.2 Hz), 2.98-3.05(m,1H), 3.15-3.23(m,2H),3.35-3.42(m,1H), 3.55(dd,1H,J=10.1 Hz,3.6 Hz), 3.90(s,3H), 3.96(s,6H),4.63(d,1H,J=15.2 Hz), 4.67(d,1H,J=15.2 Hz), 7.09(dd,1H,J=5.1 Hz,1.6 Hz),7.21(s,2H), 7.53(d,1H,J=0.6 Hz), 8.61(dd,1H,J=5.0 Hz,0.7 Hz).

EXAMPLE 8 Synthesis of(3S)-3-(2-methylpropyl)-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine2.5 hydrochloride

(3S)-3-(2-methylpropyl)-2-oxo-1-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]piperazine(100 mg) and 4-chloromethyl-2-(3,4,5-trimethoxyphenyl)pyridine (71 mg)were reacted in the same manner as in Example 4 to obtain the titlecompound as a hydrochloride.

Yield: 58 mg (50%).

¹H-NMR (measured as a free base, 400 MHz, CDCl₃) δ: 0.90(d,3H,J=6.2 Hz),0.97(d,3H,J=6.4 Hz), 1.78-1.81(m,1H), 1.90-2.00(m,2H), 2.63(dt,1H,J=13.3Hz,4.9 Hz), 3.12-3.25(m,2H), 3.30(t,1H,J=5.9 Hz), 3.37-3.42(m,1H),3.64(d,1H,J=14.3 Hz), 3.90(s,6H), 3.96(m,13H), 4.50(d,1H,J=5.0 Hz),4.86(d,1H,J=5.2 Hz), 7.12(d,1H,J=3.7 Hz), 7.21(d,1H,J=5.1 Hz),7.22(s,2H), 7.23(s,2H), 7.56(s,1H), 7.65(s,1H), 8.61(d,1H,J=4.9 Hz),8.64(d,1H,J=4.7 Hz).

m/z (EI): 670 [M⁺].

PREPARATION EXAMPLE 39 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(9-fluorenylmethoxycarbonyl)-L-phenylalanineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]amine(973 mg) and N-(9-fluorenylmethoxycarbonyl)-L-phenylalanine (871 mg)were treated in the same manner as in Preparation Example 9 to obtainthe title compound.

Yield: 1.35 g (75%).

PREPARATION EXAMPLE 40 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-L-phenylalanineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(9-fluorenylmethoxycarbonyl)-L-phenylalanineamide (1.35 g) was treated in the same manner as in Preparation Example10 to obtain the title compound.

Yield: 865 mg (89%).

PREPARATION EXAMPLE 41 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-phenylalanineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-L-phenylalanineamide (865 mg) was treated in the same manner as in Preparation Example4 to obtain the title compound.

Yield: 1.07 g (94%).

PREPARATION EXAMPLE 42 Synthesis ofN-(2-hydroxyethyl)-N-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-phenylalanineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-phenylalanineamide (1.06 g) was treated in the same manner as in Preparation Example12 to obtain the title compound.

Yield: 983 mg (theoretical amount).

PREPARATION EXAMPLE 43 Synthesis of(3S)-3-benzyl-4-(2-nitrobenzenesulfonyl)-2-oxo-1-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-piperazine

N-(2-Hydroxyethyl)-N-[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-L-phenylalanineamide (921 mg) was treated in the same manner as in Preparation Example13 to obtain the title compound. Since removal of by-products could notbe completely conducted, this compound was used in the next reactionwithout conducting further purification.

PREPARATION EXAMPLE 44 Synthesis of(3S)-3-benzyl-2-oxo-1-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]piperazine

The whole amount of the reaction mixture obtained in Preparation Example43 was treated in the same manner as in Preparation Example 6 to obtainthe title compound.

Yield: 430 mg (69% by 2 steps).

¹H-NMR (400 MHz, CDCl₁ ₃) δ: 1.62(br,1H), 2.90-2.96(m,2H),3.09-3.17(m,2H), 3.38(dt,1H,J=10.9 Hz,4.3 Hz), 3.52(dd,1H,J=13.6 Hz,3.4Hz), 3.74(dd,1H,J=9.8 Hz,3.5 Hz), 3.90(s,3H), 3.96(s,6H),4.65(d,1H,J=15.2 Hz), 4.70(d,1H,J=15.2 Hz), 7.05(dd,1H,J=5.1 Hz,1.6 Hz),7.22(s,2H), 7.25-7.34(m,5H), 7.54(s,1H), 8.61(d,1H,J=5.1 Hz).

EXAMPLE 9 Synthesis of(3S)-3-benzyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazinesesqui-hydrochloride

(3S)-3-benzyl-2-oxo-1-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]piperazine(89 mg) and 4-chloromethyl-2-(3,4,5-trimethoxyphenyl)pyridine (59 mg)were reacted in the same manner as in Example 4 to obtain the titlecompound as a hydrochloride.

Yield: 102 mg (72%).

¹H-NMR (measured as a free base, 400 MHz, CDCl₃) δ: 2.52-2.58(m,1H),3.01(dt,1H,J=12.9 Hz,4.5 Hz), 3.11-3.13(m,2H), 3.26(dd,1H,J=14.2 Hz,4.3Hz), 3.39(dd,1H,J=14.2 Hz,5.8 Hz), 3.51(d,1H,J=14.4 Hz)¤3.58(t,1H,J=4.8Hz), 3.90(s,3H), 3.91(s,3H), 3.94(s,12H), 4.13(d,1H,J=14.3 Hz),4.39(d,1H,J=15.2 Hz), 4.87(d,1H,J=15.2 Hz), 6.79(d,1H,J=4.1 Hz),7.00(d,1H,J=4.7 Hz), 7.17-7.30(m,9H), 7.48(s,1H), 7.50(s,1H),8,53(d,1H,J=5.1 Hz), 8.55(d,1H,J=5.1 Hz).

m/z (EI): 704 [M⁺].

PREPARATION EXAMPLE 45 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(9-fluorenylmethoxycarbonyl)-Nω-trityl-L-asparagineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]amine(725 mg) and N-(9-fluorenylmethoxycarbonyl)-Nω-trityl-L-asparagine (1.00g) were treated in the same manner as in Preparation Example 9 to obtainthe title compound.

Yield: 563 mg (33%).

PREPARATION EXAMPLE 46

Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nω-trityl-L-asparagineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nω-(9-fluorenylmethoxycarbonyl)-Nω-trityl-L-asparagineamide (563 mg) was treated in the same manner as in Preparation Example10 to obtain the title compound.

Yield: 396 mg (90%).

PREPARATION EXAMPLE 47 Synthesis ofN-[2-(tert-butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-Nω-trityl-L-asparagineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nω-trityl-L-asparagineamide (396 mg) was treated in the same manner as in Preparation Example4 to obtain the title compound.

Yield: 465 mg (95%).

PREPARATION EXAMPLE 48 Synthesis ofN-(2-hydroxyethyl)-N-[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-Nω-trityl-L-asparagineamide

N-[2-(tert-Butyldimethylsilyloxy)ethyl]-N-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-Nω-trityl-L-asparagineamide (410 mg) was treated in the same manner as in Preparation Example12 to obtain the title compound.

Yield: 410 mg (88%).

PREPARATION EXAMPLE 49 Synthesis of(3S)-4-(2-nitrobenzenesulfonyl)-2-oxo-1-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-3-[2-(tritylaminocarbonyl)methyl]piperazine

N-(2-Hydroxyethyl)-N-[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]-Nα-(2-nitrobenzenesulfonyl)-Nω-trityl-L-asparagineamide (410 mg) was treated in the same manner as in Preparation Example13 to obtain the title compound. Since removal of by-products could notbe completely conducted, this compound was used in the next reactionwithout conducting further purification.

PREPARATION EXAMPLE 50 Synthesis of(3S)-2-oxo-1-[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]-3-[2-(tritylaminocarbonyl)methyl]-piperazine

The whole amount of the reaction mixture obtained in Preparation Example49 was treated in the same manner as in Preparation Example 6 to obtainthe title compound.

Yield: 233 mg (75% by 2 steps).

¹H-NMR (400 MHz, CDCl₃) δ: 2.74(br,1H), 2.87-3.08(m,5H),3.35-3.39(m,1H), 3.73-3.76(m,1H), 3.89(s,3H), 3.93(s,6H),4.45(dd,1H,J=15.3 Hz,6.5 Hz), 4.72(dd,1H,J=15.3 Hz,7.1 Hz),7.03(d,1H,J=3.5 Hz), 7.18-7.28(m,18H), 7.47(s,1H), 8.53(d,1H,J=4.9 Hz).

PREPARATION EXAMPLE 51 Synthesis of(3S)-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxy-phenyl)pyridin-4-yl]methyl]-3-[2-(tritylaminocarbonyl)-methyl]piperazine

(3S)-2-Oxo-1-[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-3-[2-(tritylaminocarbonyl)methyl]piperazine(233 mg) and 4-chloromethyl-2-(3,4, 5-trimethoxyphenyl)-pyridine (104mg) were reacted in the same manner as in Example 4 to obtain the titlecompound.

Yield: 292 mg (90%).

¹H-NMR (400 MHz, CDCl₃) δ: 2.91-3.10(m,3H), 3.16(d,1H,J=13.2 Hz),3.29-3.44(m,3H), 3.51-3.59(m,1H), 3.86-3.95(m,19H), 4.40(d,1H,J=13.2Hz), 4.95(d,1H,J=15.4 Hz), 6.98(d,1H,J=4.9 Hz), 7.02(d,1H,J=4.9 Hz),7.16-7.26(m,19H), 7.45(s,1H), 7.68(s,1H), 7.72(s,1H), 8.47(d,1H,J=5.1Hz), 8.51(d,1H,J=4.9 Hz).

EXAMPLE 10 Synthesis of(3S)-3-carbamidomethyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazinedihydrochloride

(3S)-2-Oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]-3-[2-(tritylaminocarbonyl)methyl]-piperazine(292 mg) was dissolved in acetic acid (2 mL), and to the solutiontrifluoroacetic acid (4 m) was added, and the mixture was stirred at 80°C. After completion of the reaction, the reaction mixture wasconcentrated under reduced pressure, chloroform was added to theresidue, and the resultant mixture was washed with a saturated aqueoussolution of sodium hydrogencarbonate, water and saturated brine, driedover anhydrous sodium sulfate and concentrated under reduced pressure.The residue was purified by column chromatography on silica gel(chloroform:ammonia-saturated methanol=20:1) to obtain the titlecompound as a hydrochloride.

Yield: 108 mg (51%).

¹H-NMR (400 MHz, CDCl₃) δ: 2.64(t,1H,J=10.5 Hz), 2.93-3.00(m,2H),3.08(d,1H,J=11.7 Hz), 3.37-3.50(m,4H), 3.80(s,3H), 3.82(s,6H),3.83(s,3H), 3.86(s,6H), 4.02(d,1H,J=15.6 Hz), 4.09(d,1H,J=14.6 Hz),5.50(d,1H,J=15.4 Hz), 7.05(s,2H), 7.13(s,2H), 7.18(d,1H,J=4.9 Hz),7.57-7.67(m,3H), 8.53(d,1H,J=5.1 Hz), 8.82(d,1H,J=5.1 Hz).

m/z (EI) : 671 [M⁺].

PREPARATION EXAMPLE 52 Synthesis ofN-(9-fluorenylmethoxycarbonyl)-L-valyl-N-(benzyl)glycine ethyl ester

N-(9-Fluorenylmethoxycarbonyl)-L-valine (1.0 g) was dissolved in a mixedsolvent of dichloromethane (10 mL) and DMF (0.1 mL), and to the solutionoxalyl chloride (374 mg) was added dropwise at 0° C. The mixture wasstirred for 30 minutes and added dropwise into a solution ofN-(benzyl)glycine ethyl ester (587 mg) and triethylamine (477 mg) indichloromethane (10 mL) at 0° C. The resultant mixture was stirred atroom temperature for 2 hours, and the reaction mixture was concentratedunder reduced pressure. Ethyl acetate was added to the residue, and themixture was washed with water and saturated brine, dried over anhydroussodium sulfate and concentrated under reduced pressure. The residue waspurified by column chromatography on silica gel (n-hexane:ethylacetate=3:1) to obtain the title compound.

Yield: 1.37 g (91%).

PREPARATION EXAMPLE 53 Synthesis of cyclo-[N-(benzyl)glycyl-L-valyl]

N-(9-Fluorenylmethoxycarbonyl)-L-valyl-N-(benzyl)glycine ethyl ester(1.23 g) was dissolved in a 20% acetonitrile solution (12 mL) ofpiperidine, and the solution was stirred at room temperature. After 30minutes, the reaction mixture was concentrated under reduced pressure.Ethyl acetate was added to the residue, and the mixture was washed withwater and saturated brine, dried over anhydrous sodium sulfate andconcentrated under reduced pressure. The residue was purified by columnchromatography on silica gel (chloroform:methanol=50:1) to obtain thetitle compound.

Yield: 558 mg (95%).

¹H-NMR (400 MHz, CDCl₃) δ: 0.88(d,3H,J=6.63 Hz), 1.03(d,3H,J=7.2 Hz),2.42-2.49(m,1H), 3.77(d,1H,J=17.8 Hz), 3.86(d,1H,J=17.8 Hz),3.93(t,1H,J=2.9 Hz), 4.45(d,1H,J=14.3 Hz), 4.76(d,1H,J=14.4 Hz),6.79(br,1H), 7.26-7.37(m,5H).

PREPARATION EXAMPLE 54 Synthesis of(3S)-1-(benzyl)-3-isopropylpiperazine

Cyclo-[N-(benzyl)glycyl-L-valyl] (558 mg) was dissolved in THF (20 mL),and to the solution lithium aluminum hydride (430 mg) was added at 0°C., and the mixture was stirred at room temperature under an argonatmosphere. After completion of the reaction, a saturated aqueoussolution of ammonium chloride was added to the reaction mixture, and anexcessive amount of a saturated aqueous solution of sodiumhydrogencarbonate was then added to conduct extraction with ethylacetate. The resultant organic layer was dried over anhydrous sodiumsulfate and concentrated under reduced pressure, and the residue wasthen purified by column chromatography on silica gel(chloroform:methanol=30:1) to obtain the title compound.

Yield: 386 mg (78%).

¹H-NMR (400 MHz, CDCl₃) δ: 0.88(d,3H,J=6.8 Hz), 0.93(d,3H,J=6.8 Hz),1.51-1.59(m,1H), 1.78(br,2H), 1.98(dt,1H,J=11.1 Hz,3.1 Hz),2.46-2.50(m,1H), 2.72(d,1H,J=10.9 Hz), 2.85-2.89(m,2H),2.98(dt,1H,J=11.9 Hz,2.7 Hz), 3.44(d,1H,J=13.1Hz), 3.56(d,1H,J=13.1 Hz),7.23-7.31(m,5H).

PREPARATION EXAMPLE 55 Synthesis of (2S)-2-isopropylpiperazine

(3S)-1-(Benzyl)-3-isopropylpiperazine (358 mg) was dissolved in aceticacid (10 mL), and to the solution 10% palladium on carbon (40 mg) wasadded, and the mixture was then stirred at 50° C. under a hydrogenatmosphere for 4 hours. The reaction mixture was filtered, and thefiltrate was concentrated under reduced pressure. The residue waspurified by column chromatography on silica gel(chloroform:ammonia-saturated methanol=20:1) to obtain the titlecompound.

Yield: 161 mg (77%).

¹H-NMR (400 MHz, CDCl₃) δ: 0.91(d,3H,J=6.8 Hz), 0.93(d,3H,J=6.6 Hz),1.48-1.57(m,1H), 1.75(br,2H), 2.31-2.45(m,2H), 2.67-2.83(m,2H),2.90(d,1H,J=11.5 Hz), 2.99-3.02(m,2H).

EXAMPLE 11 Synthesis of(2S)-2-isopropyl-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-2-yl]methyl]piperazinetrihydrochloride

(2S)-2-Isopropylpiperazine (25 mg) and4-chloromethyl-2-(3,4,5-trimethoxyphenyl)pyridine (117 mg) were reactedin the same manner as in Example 1 to obtain the title compound as ahydrochloride.

Yield: 129 mg (theoretical amount).

¹H-NMR (400 MHz, CDCl₃) δ: 0.95(d,3H,J=6.8 Hz), 0.98(d,3H, J=6.8 Hz),1.76-1.91(m,1H), 2.14-2.34(m,4H), 2.62(d,1H,J=10.1 Hz), 2.76-2.82(m,2H),3.24(d,1H,J=14.4 Hz), 3.50(d,1H,J=14.1 Hz), 3.62(d,1H,J=14.1 Hz),3.90(s,6H), 3.96(s,6H), 3.97(s,6H), 4.17(d,1H,J=14.4 Hz),7.21-7.27(m,6H), 7.64(s,1H), 7.65(s,1H), 8.58(d,1H,J=4.7 Hz),8.59(d,1H,4.5 Hz).

m/z (EI): 642 [M⁺].

PREPARATION EXAMPLE 56 Synthesis ofN-(9-fluorenylmethoxycarbonyl)-L-leucyl-N-(benzyl)glycine ethyl ester

N-(9-Fluorenylmethoxycarbonyl)-L-leucine (1.31 g) and N-(benzyl)glycineethyl ester (738 mg) were reacted in the same manner as in PreparationExample 52 to obtain the title compound.

Yield: 1.65 g (84%).

PREPARATION EXAMPLE 57 Synthesis of cyclo-[N-(benzyl)glycyl-L-leucyl]

N-(9-Fluorenylmethoxycarbonyl)-L-leucyl-N-(benzyl)glycine ethyl ester(1.79 g) was reacted in the same manner as in Preparation Example 53 toobtain the title compound.

Yield: 775 mg (88%).

¹H-NMR (400 MHz, CDCl₃) δ: 0.95(d,3H,J=6.5 Hz), 0.98(d,3H,J=6.5 Hz),1.62-1.67(m,1H), 1.75-1.85(m,2H), 3.80(d,1H,J=17.4 Hz), 3.86(d,1H,J=17.2Hz), ¤04.04(dt,1H, J=6.1 Hz,3.2 Hz), 4.54(d,1H,J=14.3 Hz),4.65(d,1H,J=14.4 Hz), 6.80(br,1H), 7. 24-7.37(m, 5H)

PREPARATION EXAMPLE 58 Synthesis of(3S)-1-(benzyl)-3-(2-methylpropyl)piperazine

Cyclo-[N-(benzyl)glycyl-L-leucyl] (775 mg) was treated in the samemanner as in Preparation Example 54 to obtain the title compound.

Yield: 700 mg (theoretical amount).

¹H-NMR (400MHz, CDCl₃) δ: 0.87(d,3H,J=6.4 Hz), 0.89(d,3H,J=6.6 Hz),1.08-1.26(m,2H), 1.61-1.71(m,3H), 2.00(dt,1H,J=11.1 Hz,3.6 Hz),2.73-2.96(m,5H), 3.39-3.58(m,2H), 7.23-7.31(m,5H).

PREPARATION EXAMPLE 59 Synthesis of (2S)-2-(2-methylpropyl)piperazine

(3S)-1-(Benzyl)-3-(2-methylpropyl)piperazine (700 mg) was treated in thesame manner as in Preparation Example 55 to obtain the title compound.

Yield: 308 mg (72%).

¹H-NMR (400 MHz, CDCl₃) δ: 0.89(d,3H,J=6.6 Hz), 0.91(d,3H,J=6.6 Hz),1.10-1.23(m,2H), 1.56(br,2H), 1.60-1.69(m,1H), 2.34(dd,1H,J=11.8 Hz,9.9Hz), 2.64-2.98(m,6H).

EXAMPLE 12 Synthesis of(2S)-2-(2-methylpropyl)-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-2-yl]methyl]piperazinetrihydrochloride

(2S)-2-(2-Methylpropyl)piperazine (28 mg) and4-chloromethyl-2-(3,4,5-trimethoxyphenyl)pyridine (117 mg) were reactedin the same manner as in Example 1 to obtain the title compound as ahydrochloride.

Yield: 129 mg (99%).

¹H-NMR (400 MHz, CDCl₃) δ: 0.86(d,3H,J=6.1 Hz) 0.90(d,3H,J=6.1 Hz),1.50-1.56(m,3H), 2.25-2.78(m,7H), 3.39(d,1H,J=14.1 Hz), 3.51(d,1H,J=14.1Hz), 3.62(d,1H,J=14.1 Hz), 3.90(s,6H), 3.97(br,13H), 7.22-7.27(m,6H),7.65(s,2H), 8.59(d,1H,J=5.3 Hz), 8.60(d,1H,J=5.5 Hz).

m/z (EI): 656 [M⁺].

PREPARATION EXAMPLE 60 Synthesis ofN-(9-fluorenylmethoxycarbonyl)-L-isoleucyl-N-(benzyl)glycine ethyl ester

N-(9-Fluorenylmethoxycarbonyl)-L-isoleucine (1.36 g) andN-(benzyl)glycine ethyl ester (770 mg) were reacted in the same manneras in Preparation Example 52 to obtain the title compound.

Yield: 1.73 g (85%).

PREPARATION EXAMPLE 61 Synthesis of cyclo-[N-(benzyl)glycyl-L-isoleucyl]

N-(9-Fluorenylmethoxycarbonyl)-L-isoleucyl-N-(benzyl)glycine ethyl ester(1.63 g) was reacted in the same manner as in Preparation Example 53 toobtain the title compound.

Yield: 973 mg (including impurities).

¹H-NMR (400 MHz, CDCl₃) δ: 0.90 (t,3H,J=7.4 Hz), 1.00(d,3H,J=7.2 Hz),1.14-1.25(m,1H), 1.35-1.43(m,1H), 2.09-2.15(m,1H), 3.77(d,1H,J=18.0 Hz),3.85(d,1H,J=17.8 Hz), 3.95-3.97(m,1H), 4.51(d,1H,J=14.3 Hz),4.69(d,1H,J=14.4 Hz), 7.25-7.36(m,6H).

PREPARATION EXAMPLE 62 Synthesis of(3S)-1-(benzyl)-3-(1-methylpropyl)piperazine

Cyclo-[N-(benzyl)glycyl-L-isoleucyl] (973 mg, including impurities) wastreated in the same manner as in Preparation Example 54 to obtain thetitle compound.

Yield: 506 mg (71% by 2 steps).

¹H-NMR (400 MHz, CDCl₃) δ: 0.87(d,3H,J=6.8 Hz), 0.87(t,3H,J=7.4 Hz),1.13-1.20(m,1H), 1.30-1.41(m,1H), 1.46-1.70(m,2H), 1.78(t,1H,J=10.4 Hz),1.97(dd,1H,J=11.1 Hz,3.3 Hz), 2.57-2.62(m,1H), 2.68-2.76(m,1H),2.82-2.89(m,2H), 2.97(dt,1H,J=11.9 Hz,2.7 Hz), 3.44(d,1H,J=13.1 Hz),3.56(d,1H,J=13.1 Hz), 7.24-7.31(5H).

PREPARATION EXAMPLE 63 Synthesis of (2S)-2-(1-methylpropyl)piperazine

(3S)-1-(Benzyl)-3-(1-methylpropyl)piperazine (506 mg) was treated in thesame manner as in Preparation Example 55 to obtain the title compound.

Yield: 202 mg (65%).

¹H-NMR (400 MHz, CDCl₃) δ: 0.87 (d,3H,J=6.8 Hz), 0.89(t,3H, J=7.4 Hz),1.12-1.23(m,1H), 1.29-1.32(m,1H), 1.44-1.52(m,1H), 1.64(br,2H),2.40-2.48(m,2H), 2.69(dt,1H,J=11.3 Hz,2.9 Hz), 2.80(dt,1H,J=11.3 Hz,2.7Hz), 2.89(d,1H,J=11.5 Hz), 2.94-3.01(m,2H).

EXAMPLE 13 Synthesis of(2S)-2-(1-methylpropyl)-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-2-yl]methyl]piperazinetrihydrochloride

(2S)-2-(1-Methylpropyl)piperazine (28 mg) and4-chloromethyl-2-(3,4,5-trimethoxyphenyl)pyridine (117 mg) were reactedin the same manner as in Example 1 to obtain the title compound as ahydrochloride.

Yield: 117 mg (90%).

¹H-NMR (400 MHz, CDCl₃) δ: 0.93 (t,3H,J=7.8 Hz), 0.95(d,3H,J=6.6 Hz),1.16-1.25(m,1H), 1.35-1.41(m,1H), 1.90-2.05(m,1H), 2.11-2.32(m,3H),2.45(br,1H), 2.65(d,1H,J=10.3 Hz), 2.74-2.79(m,2H), 3.17(d,1H,J=14.3Hz), 3.52(d,1H,J=14.1 Hz), 3.60(d,1H,J=14.1 Hz), 3.90(s,6H), 3.96(s,6H),3.97(s,6H), 4.19(d,1H,J=14.2 Hz), 7.21-7.27(m,6H), 7.65(s,2H),8.58(d,1H,J=3.1 Hz), 8.60(d,1H,J=3.1 Hz).

m/z (EI): 656 [M⁺]

Test Example 1

(Inhibitory Effect on Cell Adhesion)

This test was conducted by reference to the method of Ross et al. (J.Biol. Chem., 267, 8537-8543 (1992)). More specifically, after humanumbilical venous endothelial cells (HUVEC) were cultured on a 48-wellplate to confluent growth, TNFα was added thereto. Upon elapsed time of5 hours after the addition, U937, which is a human monocytic/histocyticcell fluorescence-labeled with PKH2 (product of Dainippon PharmaceuticalCo., Ltd.), was added in a proportion of 1×10⁶ cells per well. After theplate was left at rest at room temperature for 1 hour, unadhered U937was washed out and lysed in 1% Triton X-100 to measure a remainingfluorescence intensity (excitation wavelength: 480 nm; measuringwavelength: 530 nm). HUVEC and U937 were cultured in EGM-2 (product ofSanko Junyaku K.K.) and 10% FCS-containing RPMI1640, respectively. Eachtest agent was added to HUVEC upon the addition of TNFα and to U937 24hours prior to the cell adhesion test. The inhibitory activity wascalculated out according to the equation [100−(C−B)/(A−B)×100 (%)],wherein A is the number of U937 cells adhered to HUVEC stimulated byTNFα when no test agent was added, B is the number of U937 cells adheredto HUVEC not stimulated by TNFα when no test agent was added, and C isthe number of U937 cells adhered to HUVEC stimulated by TNFα when thetest agent was added. The results are shown in Table 1. As controlcompounds, Test Compound 1 described in Japanese Patent ApplicationLaid-Open No. 9-143075 and dilazep described in Japanese PatentApplication Laid-Open No. 11-92382 were simultaneously evaluated.

TABLE 1 Inhibitory activity of each compound at 1, 10 μM against celladhesion Percent inhibition (%) Example 1 μM 10 μM 1 76 78 2 42 64 3 3490 4 60 79 5 51 85 6 63 77 7 51 70 8 63 79 9 38 86 Test compound 1  5 51Dilazep 12 25

Specific formulation examples will hereinafter be described.

PREPARATION EXAMPLE 64 Capsule Preparation

(3S)-3-Isopropyl-2-oxo-N,N′-bis[[2-(3,4,5- 30 mgtrimethoxyphenyl)pyridin-4-yl]methyl]- piperazine sesquihydrochlorideMicrocrystalline cellulose 30 mg Lactose 30 mg Magnesium stearate 3 mgTotal amount 93 mg.

The above ingredients were mixed in accordance with a method known perse in the art and then charged in a gelatin capsule to obtain a capsulepreparation.

PREPARATION EXAMPLE 65 Table Preparation

(3S)-3-Isopropyl-2-oxo-N,N′-bis[[2-(3,4,5- 30 mgtrimethoxyphenyl)pyridin-4-yl]methyl]- piperazine sesquihydrochlorideStarch 44 mg Starch (for glue) 5.6 mg Magnesium stearate 0.4 mg Calciumcarboxymethyl cellulose 20 mg Total amount 100 mg.

The above ingredients were mixed in accordance with a method known perse in the art to obtain a tablet preparation.

PREPARATION EXAMPLE 66 Injection Preparation

(3S)-3-Isopropyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazinesesqui- hydrochloride (100 mg) and sodium chloride (900 mg) weredissolved in distilled water (about 80 mL) for injection, and distilledwater for injection was added to the resultant solution to 100 mL intotal. This diluted solution was sterilized by filtration and thensubdivided and charged into 10 light-screening ampoules, and theampoules were sealed to obtain sterile injection preparations.

As described above, the compounds (1) according to the present inventionhave excellent inhibitory effects on both cell adhesion and cellinfiltration and are useful for prevention or treatment of diseases suchas allergy, asthma, rheumatism, arteriosclerosis and inflammation.

Obviously, numerous modifications of the above teachings are apparent tothose skilled in the art. Therefore, within the scope of the appendedclaims the invention can be practiced otherwise than as specificallydescribed herein.

What is claimed is:
 1. A piperazine compound of formula (1):

wherein X is —CH₂—, —C(O)— or —CH(CH₃)—; R¹ is a hydrogen atom or alkylgroup; and R² is a hydrogen atom, alkyl group, hydroxyalkyl group,arylalkyl group, heteroarylalkyl group, carboxyalkyl group,carboxamidoalkyl group, aminoalkyl group or guanidinoalkyl group; anacid-addition salt thereof, or a hydrate thereof.
 2. The piperazinecompound of claim 1, wherein R¹ is a hydrogen atom or C₁-C₆-alkyl group;and R² is a hydrogen atom, C₁-C₆-alkyl group, hydroxy-C₁-C₆-alkyl group,C₆-C₁₀-aryl-C₁-C₆-alkyl group, 5- or 6-membered heteroaryl-C₁-C₆-alkylgroup having 1 or 2 nitrogen atoms, carboxy-C₁-C₆-alkyl group,carboxamido-C₁-C₆-alkyl group, amino-C₁-C₆-alkyl group orguanidino-C₁-C₆-alkyl group.
 3. A pharmaceutical composition comprisingas an active ingredient, a piperazine compound of formula (1):

wherein X is —CH₂—, —C(O)— or —CH(CH₃)—; R¹ is a hydrogen atom or alkylgroup; and R² is a hydrogen atom, alkyl group, hydroxyalkyl group,arylalkyl group, heteroarylalkyl group, carboxyalkyl group,carboxamidoalkyl group, aminoalkyl group or guanidinoalkyl group; anacid-addition salt thereof, or a hydrate thereof, and a pharmaceuticallyacceptable carrier.
 4. The composition of claim 3, wherein R¹ is ahydrogen atom or C₁-C₆-alkyl group; and R² is a hydrogen atom,C₁-C₆-alkyl group, hydroxy-C₁-C₆-alkyl group, C₆-C₁₀-aryl-C₁-C₆-alkylgroup, 5- or 6-membered heteroaryl-C₁-C₆-alkyl group having 1 or 2nitrogen atoms, carboxy-C₁-C₆-alkyl group, carboxamido-C₁-C₆-alkylgroup, amino-C₁-C₆-alkyl group or guanidino-C₁-C₆-alkyl group.
 5. Thecomposition of claim 3, comprising an effective amount of the piperazinecompound for treating a disease caused by cell adhesion and/or cellinfiltration.
 6. The composition of claim 5, wherein the disease isselected from the group consisting of allergy, asthma, inflammation,rheumatism and arteriosclerosis.
 7. A method for treating a diseaseselected from the group consisting of allergy, asthma, inflammation,rheumatism, autoimmune disease and arteriosclerosis, which comprisesadministering to a patient in need thereof a piperazine compound offormula (1)

wherein X is —CH₂—, —C(O)— or —CH(CH₃)—; R¹ is a hydrogen atom or alkylgroup, and R² is a hydrogen atom, alkyl group, hydroxyalkyl group,arylalkyl group, heteroarylalkyl group, carboxyalkyl group,carboxamidoalkyl group, aminoalkyl group or guanidinoalkyl group; anacid-addition salt thereof, or a hydrate thereof.
 8. The method of claim7, wherein R¹ is a hydrogen atom or C₁-C₆-alkyl group; and R² is ahydrogen atom, C₁-C₆-alkyl group, hydroxy-C₁-C₆-alkyl group,C₆-C₁₀-aryl-C₁-C₆-alkyl group, 5- or 6-membered heteroaryl-C₁-C₆-alkylgroup having 1 or 2 nitrogen atoms, carboxy-C₁-C₆-alkyl group,carboxamido-C₁-C₆-alkyl group, amino-C₁-C₆-alkyl group orguanidino-C₁-C₆-alkyl group.
 9. The method of claim 7, wherein saiddisease is asthma and wherein said disease comprises infiltration ofeosinophils into a bronchus.
 10. The method of claim 7, wherein saiddisease is arteriosclerosis and wherein said disease comprisesinfiltration of macrophages and T lymphocytes into the aorta.
 11. Themethod of claim 7, wherein said disease is inflammation and wherein saiddisease comprises infiltration of T lymphocytes and eosinophils into theskin.
 12. The method of claim 7, wherein said disease is rheumatism andwherein said disease comprises infiltration of leukocytes intorheumatoid synovial tissue.
 13. The method of claim 7, wherein saiddisease is allergy, said allergy disease selected from the groupconsisting of bronchial asthma, dermatitis, rhinitis and conjunctivitis.14. The method of claim 7, wherein said disease is an autoimmune diseaseselected from the group consisting of rheumatoid arthritis, nephritis,inflammatory bowel disease, diabetes, and chronic inflammatory disease.15. The method of claim 7, wherein said disease is inflammation of thecolon.
 16. The method of claim 7, wherein said piperazine compound isadministered in a dose of from 1 to 1,0000 mg per day.
 17. The method ofclaim 7, wherein said piperazine compound is administered orally. 18.The method of claim 7, wherein said piperazine compound is administeredparenterally.
 19. The method of claim 7, wherein said piperazinecompound is selected from the group consisting of(3S)-3-Isopropyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-piperazinesesquihydrochloride,cis-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-2,6-dimethylpiperazinetetrahydrochloride,trans-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]-2,5-dimethylpiperazinetetrahydrochloride,N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-2-hydroxymethylpiperazinedimaleate,2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]piperazine2.5 hydrochloride,(3S)-3-methyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine2.5 hydrochloride,(3R)-3-isopropyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazinetrihydrochloride,(3S)-3-(2-methylpropyl)-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine2.5 hydrochloride, and(3S)-3-benzyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl]pyridin-4-yl]methyl]piperazinesesquihydrochloride.
 20. The piperazine compound of claim 1, which isselected from the group consisting of(3S)-3-Isopropyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-piperazinesesquihydrochloride,cis-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-2,6-dimethylpiperazinetetrahydrochloride,trans-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]-2,5-dimethylpiperazinetetrahydrochloride,N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-2-hydroxymethylpiperazinedimaleate,2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]piperazine2.5 hydrochloride,(3S)-3-methyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine2.5 hydrochloride,(3R)-3-isopropyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazinetrihydrochloride,(3S)-3-(2-methylpropyl)-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine2.5 hydrochloride, and(3S)-3-benzyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl]pyridin-4-yl]methyl]piperazinesesqui-hydrochloride.
 21. The composition of claim 3, wherein saidpiperazine compound is selected from the group consisting of(3S)-3-Isopropyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-piperazinesesquihydrochloride,cis-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-2,6-dimethylpiperazinetetrahydrochloride,trans-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]-2,5-dimethylpiperazinetetrahydrochloride,N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]-2-hydroxymethylpiperazinedimaleate,2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)-pyridin-4-yl]methyl]piperazine2.5 hydrochloride,(3S)-3-methyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine2.5 hydrochloride, (3R)-3-isopropyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine trihydrochloride,(3S)-3 -(2-methylpropyl)-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl)pyridin-4-yl]methyl]piperazine 2.5 hydrochloride,and(3S)-3-benzyl-2-oxo-N,N′-bis[[2-(3,4,5-trimethoxyphenyl]pyridin-4-yl]methyl]piperazinesesqui-hydrochloride.